Gonapeptyl

During IVF trials, stimulation is achieved with 100 to 300 IU of gonadotrophins depending on age, body mass index (BMI) and the Anti-Mullerian hormone (AMH) level (FSHrec: Puregon,; MSD, Oss, the Netherlands; GonalF: Merck-Serono, Geneva, Switzerland) or human Menopausal Gonadotrophin (Menopur, Ferring, Alost, Belgique). Ovulation was controlled by either gonadotropin-releasing hormone (GnRH) antagonist (Cetrorelix, Merck Serono or Ganirelix, MSD) or GnRH agonist (Buserelin: Suprefact; Sanofi-Avantis, Diegem, Belgium or triptoréline: Gonapeptyl, Ferring) respectively in 68% (170 cycles) and 32% (81 cycles) of cases. The GnRH antagonist is administered according to a flexible protocol which is once daily, when at least one follicle has reached a diameter of 14 mm and / or the oestradiol (E2) level is at 400 pg / ml. When a GnHR agonist is used, the long protocol is applied in the majority of such cycles. Triggering was obtained with 5000 IU of human chorionic gonadotrophin (hCG) (Pregnyl, MSD, Bruxelles, Belgium).

Controlled ovarian stimulation (COS) was performed using recombinant follicle stimulating hormone (Gonal-F®; recFSH, Merck Serono, Switzerland) and/or highly purified urinary gonadotropins (Menopur®; hpHMG, Ferring, Denmark). The gonadotropin starting dose was individualized to patient characteristics and dose adjustments performed according to transvaginal ultrasound findings and estradiol serum levels. A GnRH antagonist (Cetrotide®; Cetrorelix, Merck Serono, Switzerland) was utilized for pituitary suppression, 0.25 mg subcutaneously daily, when at least one follicle attained 12–14 mm in diameter. Final oocyte maturation was triggered using a GnRH agonist (Gonapeptyl®; triptorelin, Ferring, Denmark). Transvaginal oocyte retrieval was performed 36 h after GnRH agonist 0.3 mg subcutaneous administration. Assisted fertilization was performed using intracytoplasmic sperm injection (ICSI). Normally fertilized zygotes were cultured in extended culture medium in a time lapse incubator (Embryoscope®; Vitrolife, Sweden). Blastocysts were considered suitable for TE biopsy and vitrification based on a morphokinetic assessment that combines the criteria of Gardner and Schoolcraft (Gardner et al., 2000) with the KID score.

Menopausal gonadotropins were used in monotherapy as described elsewhere^{37 (link)}. All women were treated with a long agonist protocol starting with oral contraceptives (OCs) (Rigevidon, Richter Gedeon, Warsaw, Poland) from the 2nd to the 5th day of the cycle. Triptorelin acetate 0.1 mg (Gonapeptyl, Ferring, Saint-Prex, Switzerland) was administered 14 days after the initiation of the OCs. Fourteen days later (7 days after the end of OC administration), urinary gonadotropins (Menopur, Ferring, Saint-Prex, Switzerland) were administered for ovarian stimulation. Follicular growth was monitored on day 8 of COH using transvaginal ultrasound, and assays evaluating serum estradiol (E2), progesterone (P), and luteinizing hormone (LH) levels were performed until pituitary ovarian downregulation was reached (i.e., E2 concentration < 50 pg/ml). Follicular growth was stimulated by administering FSH (Menopur, Ferring, Saint-Prex, Switzerland), considering individual endocrine response as well as ovarian reaction estimated as the presence of at least an 18-mm-diameter follicle. Oocyte retrieval under the guidance of transvaginal ultrasound was performed 36–38 h after ovulation induction with human chorionic gonadotropin (hCG) injection.

COH was performed with the GnRH antagonist protocol. Recombinant FSH (150–300 IU, Gonal-F; Serono) and/or hMG (75–150) IU; (Merional; IBSA) was administered on day 2 of the menstrual period. Starting on the sixth day of controlled ovarian stimulation, the ovarian response was monitored by serial transvaginal ultrasound (TV-USG) and by measuring serum E₂ and P₄ levels. When the leading follicle exceeded 13 mm in diameter, 0.25 mg of GnRH antagonist (Cetrotide; Serono) was started daily until the day of the last trigger. When at least two follicles reached 18 mm in diameter, patients were administered 250 μg of human chorionic gonadotropin (hCG; Ovitrelle, Serono) or 0.2 mg of triptorelin (Gonapeptyl, Ferring), and oocyte retrieval was scheduled 35 hours after the trigger administration [16 (link)].
The oocyte retrieval, denudation, and ICSI procedures were performed as described previously by Serdarogullari et al. [17 ]. ICSI was the fertilization method in all of the cycles included in this study. After microinjection, oocytes were cultured individually in a special pre-equilibrated culture dish. In our study, single-step media, namely, Continuous Single Culture Complete (CSCM-C) with Human Serum Albumin (Irvine Scientific) was used for embryo culture throughout the culture period.

The gonadotropin-releasing hormone (GnRH) antagonist protocol was the preferred method for ovarian stimulation. On the 2^nd or 3^rd day of the menstrual cycle, gonadotrophin injections were started by using recombinant follicle-stimulating hormone (Gonal-F; Merck Serono, Geneva, Switzerland) and/or highly purified human menopause gonadotrophins (hp-hMG) (75-150 IU, Merional; IBSA) preparations. The dose regimens were designated at the physician’s preference. When the leading follicle exceeded 13 mm in diameter, 0.25 mg of GnRH antagonist (Cetrotide; Serono) was started daily until the day of maturation trigger. Maturation of the oocytes was induced either with the use of 250 µg of human chorionic gonadotropin (hCG; Ovitrelle, Serono) or 0.2 mg triptorelin (Gonapeptyl, Ferring). Transvaginal sonography (TV-USG)-guided oocyte retrieval was performed 35-36 hours later.

The women received an initial dose of 300IU menotropin (Merional^®) for four days, and on the fifth day of induction, the dose was reduced to 150IU/day. The first ultrasound to assess follicular development was performed between the fifth and the sixth day of induction. After identifying 02 follicles ≥14mm or 01 ≥16mm, 01 ampoule/day of ganirelix (Orgalutran^®, Merck Sharp & Dohme B.V., Netherlands) was started up to 24 hours before the trigger. In the presence of two or more follicles with an average diameter of 18mm, a trigger with triptorelin (Gonapeptyl^®, Ferring GmbH, Kiel, Germany) was prescribed and the egg collection was performed after 34-36 hours. After 2-4 hours of ovarian puncture, ICSI was performed, and the embryos were vitrified on the third day (D3) or on the blastocyst stage.